A routine system for generation of transgenic rice (Oryza sativa L.) plants by microprojectile bombardment of embryogenic cell clusters
We have developed a routine system, based on microprojectile bombardment of embryogenic cell clusters for recovery of fertile transgenic rice (Oryza sativa L.) plants. Embryogenic cell clusters were established within 3–4 months from calli derived from mature seeds of nine diverse cultivars. The cell clusters were bombarded with plasmids carrying genes encoding hygromycin phosphotransferase (hph, conferring resistance to hygromycin B) and β-glucuronidase (gus A), both driven by the cauliflower mosaic virus (CaMV)35S promoter. Resistant calli were readily recovered from hygromycin-containing medium. On average 4.6 resistant calli were recovered from each bombarded plate. Transgenic plants were routinely regenerated from such resistant calli. In this system, where the transformation procedure was based on bombardment of embryogenic cell clusters, there were no escapes. Data from Southern blot analysis of T0 and T1 plants confirmed that the transgene was stably integrated into the host genome and inherited in a Mendelian fashion.
Ghosh Biswas, G.C.; Chen DongFang; Elliott, M.C. A routine system for generation of transgenic rice (Oryza sativa L.) plants by microprojectile bombardment of embryogenic cell clusters. Plant Science (1998) 133 (2) 203-210. [DOI: 10.1016/S0168-9452(98)00013-2]